tbuckley/cabot-search huggingface dataset:
OralsO01 Loss of TGFBR2 and activation of PI3 kinase signalling is associated with metastatic cutaneous squamous cell carcinomas
Authors: Mairi Treanor-Taylor, Ying Zhang, Daniel  Schreyer, Catherine Harwood, Irene  Leigh, Peter Bailey, David Wotton, Gareth Inman
Journal: British Journal of Dermatology (2023)

Abstract Cutaneous squamous cell carcinoma (cSCC) can be a relatively low-risk cancer type as it is typically curable upon surgical resection; however, in advanced cases, treatment options are lacking and patient outcomes are poor. The molecular mechanisms that drive metastasis and the associated targets for therapeutic intervention are yet to be determined. To investigate drivers of metastatic cSCC, we undertook whole-genome sequencing of matched primary and metastatic cSCC samples from two patients. Analysis of single nucleotide variants and copy number changes revealed the high tumour mutational burden of this cancer type due to ultraviolet-induced DNA damage, highlighting the challenge faced in filtering out passenger mutations and identifying oncogenic events. Variations unique to the metastases were obtained, and their pathogenicity was assessed. Damaging mutations were found in transforming growth factor beta receptor 2 (TGFBR2) and genes in the PTEN/PI3K/Akt signalling pathway, specifically PTEN, PIK3R1 and PIK3CA, which have previously occurred in cancer samples included in COSMIC, cBioPortal, Cancer Hotspots and Genomic Data Commons. To validate functionally the role of TGFBR2 and PTEN/PI3K/Akt pathway mutations in skin cancers, we generated conditional, inducible Ptenfl/fl;K14Cre+, Pten+/fl;Tgfbr2fl/fl;K14Cre+ and Ptenfl/fl;Tgfbr2fl/fl; K14Cre+ mice. Loss of both Tgfbr2 and Pten in mice was associated with increased proliferation (P = 2.1 × 106), invasion, lymph node metastases and poorer tumour-free (P &amp;lt; 9.0 × 107) and overall survival (P &amp;lt; 2.0 × 106). These data showcase the role of transforming growth factor-β and PTEN/PI3K/Akt signalling pathways on the development of metastatic cSCC.
P09 Short-term rho-associated kinase inhibitor treatment promotes primary keratinocyte growth without affecting keratinocyte stem cell properties
Authors: Vignesh Jayarajan, George T Hall, Theodoros  Xenakis, Neil Bulstrode, Dale  Moulding, Sergi Castellano, Wei-Li Di
Journal: British Journal of Dermatology (2023)

Abstract Keratinocyte stem cells (KSCs) can be cultured as ex vivo epidermal sheets and are attractive for cell and gene therapy. However, the growth and expansion of primary keratinocytes (including KSCs) in in vitro culture can be inefficient as KSCs undergo apoptotic cell death, known as anoikis when isolated from skin biopsies and dissociated from their extracellular environment. This further affects the survival and stemness of KSCs in in vitro culture and hinders clinical applications. Rho-associated kinase inhibitor (ROCKi) has been used to overcome this obstacle. However, it risks changing the characteristics of stem cells. In this study, we studied the effect of the Y-27632 ROCKi on cultured primary human keratinocytes. Following a short-term (6 days) ROCKi treatment, we assessed keratinocyte proliferation and differentiation potential and KSC stemness at the molecular-, cellular- and single-cell levels. A significant increase in colony-forming efficiency associated with increased expression of proliferation markers and mitochondrial mass was observed in treated cells vs. untreated cells. Similarly, treated cells showed a distinct transcriptional profile at the single-cell level, but these changes became indistinguishable after discontinuation of ROCKi treatment. Single-cell transcriptome analysis further confirmed that short-term ROCKi treatment did not affect stem cell characteristics. Finally, we showed that ROCKi rapidly activated Akt and upstream of ERK signalling pathways in primary human keratinocytes. In conclusion, rapid proliferation due to short-term ROCKi treatment was transient and reversible, and did not affect the KSC population or their characteristics. We thus present a safer approach for the use of ROCKi in ex vivo clinical applications.
O16 The novel roles of laminin α3 in epidermal barrier function and metabolism
Authors: Piotr Parzymies, Eleri Mai Jones, Andrew  Mayes, David Messenger, Edel O’Toole, Matthew  Caley
Journal: British Journal of Dermatology (2023)

Abstract The skin serves an important barrier function performed by epidermal lipids, cornified epithelial cells and cell–cell adhesions. This function is compromised in old age and exhibits certain abnormalities such as flattened dermal–epidermal junction or improper lipid metabolism, and is also associated with mitochondrial dysfunction. The levels of a basement membrane protein laminin 332 have been shown to decrease as the skin ages. We have previously demonstrated that loss of laminin 332 leads to an upregulation of genes involved in cholesterol biosynthesis and is associated with an altered skin lipid profile. We showed that this change is caused by an abnormal intracellular cholesterol transport that is actin-dependent. To further elucidate the mechanism around cholesterol transport, we have identified five proteins of interest for further investigation. We have shown that two of those proteins, NPC2 and Myo5b, are decreased with the loss of the α3 laminin 332 subunit in laminin α3 knockdown nTERT keratinocytes vs. shC controls. Interestingly, the NPC2 binding partner, NPC1, appears to be increased in the laminin α3 knockdown nTERT keratinocytes. Our analysis has also demonstrated that e-cadherin, a crucial cell–cell adhesion protein, as well as several differentiation markers, are altered with the loss of laminin α3. We have carried out laminin 332 recovery experiments in two (2D) and three dimensions, which showed that the addition of recombinant laminin 332 restores the proper levels of myo5b and e-cadherin, suggesting a potential therapeutic effect. We have also performed proteomic, lipidomic, and transcriptomic studies in 2D, which demonstrated statistically significant changes (Q-value ≤ 0.01) in proteins involved in actin and microtubule regulation, cell–cell adhesion and showed unexpected mitochondrial metabolic changes associated with the loss of laminin α3. These findings show that laminin α3 plays numerous, previously unreported roles in epidermal lipid transport, cell–cell adhesion, differentiation and metabolism, and suggests a potential therapeutic role of the protein in skin ageing.
Malignancy risk in patients with atopic dermatitis: a population-based cohort study
Authors: Joy Wan, Daniel B Shin, Maha N Syed, Katrina Abuabara, Adina R Lemeshow, Zelma C Chiesa Fuxench, Joel M Gelfand
Journal: British Journal of Dermatology (2023)

Abstract Background Atopic dermatitis (AD) is associated with immunological dysfunction, which may influence cancer development. Previous studies of AD and cancer demonstrate inconsistent results and few of these studies examined children or AD severity and treatment. Objectives To determine malignancy risk among children and adults with AD. Methods We conducted a cohort study using electronic health records data from UK general practices in The Health Improvement Network between 1994 and 2015. Children (&amp;lt; 18 years old) and adults (≥ 18 years old) with AD were matched on age, practice and index date to patients without AD. AD was categorized as mild, moderate or severe using treatments and dermatology referrals as proxies. The primary outcome was any incident malignancy, including in situ malignancy, identified using diagnosis codes and categorized into haematological, skin and solid organ malignancies. Secondary outcomes included specific malignancies: leukaemia, lymphoma, melanoma, nonmelanoma skin cancer (NMSC) and common solid-organ cancers. Results Among 409 431 children with AD (93.2% mild, 5.5% moderate, 1.3% severe) and 1 809 029 children without AD who had median follow-up of 5–7 years, the incidence rates of malignancy were 1.9–3.4 and 2.0 per 10 000 person-years (PY), respectively. The adjusted risk of malignancy overall did not differ with respect to AD [hazard ratio (HR) 1.02 (95% confidence interval 0.92–1.12)]. Severe AD was associated with increased lymphoma risk [HR 3.18 (1.41–7.16), excluding cutaneous T-cell lymphoma (CTCL)], and mild AD was associated with increased NMSC risk [1.55 (1.06–2.27)]. Among 625 083 adults with AD (65.7% mild, 31.4% moderate, 2.9% severe) and 2 678 888 adults without AD who had median follow-up of 5 years, incidence rates of malignancy were 97.4–125.3 per 10 000 PY and 103.7 per 10 000 PY, respectively. The adjusted risk of any malignancy did not differ with respect to AD [HR 1.00 (0.99–1.02)]. However, adults with severe AD had a twofold higher risk of non-CTCL lymphoma. AD was also associated with slightly higher skin cancer risk [HR 1.06 (1.04–1.08)] and slightly lower solid cancer risk [0.97 (0.96–0.98)] but results varied by specific cancers and AD severity. Conclusions Epidemiological evidence does not support a strong overall malignancy risk in AD but lymphoma risk may be increased with severe AD.
P32 Restoring the ageing skin barrier <i>in vitro</i>
Authors: Priya Garcha, Eleri Jones, Jennifer Shelley, Matthew Caley
Journal: British Journal of Dermatology (2023)

Abstract The skin provides an essential barrier to the body. As we age, the make-up of this barrier changes, with a reduction of stratum corneum lipids. This loss of lipids as we age can lead to the formation of dry and easily damaged skin, allowing for water loss from the skin surface. One mechanism for this reduction in stratum corneum lipids is dysfunctional lipid transport. To investigate lipid transport within the skin we have developed a two-dimensional high-throughput cholesterol transport assay. Using this assay we have screened 200 naturally occurring compounds and 800 plant extracts to determine their ability to improve cholesterol transport. We identified seven compounds that were able to increase cholesterol transport with no cellular toxicity. Three compounds were selected for further testing in three-dimensional (3D) organotypic skin models with or without defects in cholesterol transport and stratum corneum lipids. Three-dimensional organotypic skin models were grown for 7 days before daily treatment with either dimethyl sulfoxide or 1 µmol L–1 of the selected compound for an additional 7 days. Immunofluorescent staining was conducted to assess skin differentiation (K14, K10, TGM1), proliferation (Ki67), basement membrane (laminin α3) and stratum corneum lipids (Nile red). We have developed a 2D screening and 3D testing platform to assess compounds and drugs for their effect on cholesterol transport. Compounds that can target cholesterol transport within keratinocytes could be a beneficial treatment to help improve the ageing skin barrier, improving dry and damaged skin.
A commentary on ‘Incidence and cost of vertebral fracture in urban China: a five-year population-based cohort study’
Authors: Lan Shen, Kun Luo, Xuelin Deng, Jingjing Liu
Journal: International Journal of Surgery (2023)

aDepartment of Emergency Medicine, Southern Central Hospital of Yunnan Province (First People's Hospital of Honghe State), Yunan, 661199, China bDepartment of Spinal Surgery, Southern Central Hospital of Yunnan Province (First People's Hospital of Honghe State), Yunan, 661199, China Sponsorships or competing interests that may be relevant to content are disclosed at the end of this article. Published online ■ ■ *Corresponding author. Address: CHINA. E-mail address: [email protected] (J. Liu). This is an open access article distributed under the Creative Commons Attribution-ShareAlike License 4.0, which allows others to remix, tweak, and build upon the work, even for commercial purposes, as long as the author is credited and the new creations are licensed under the identical terms. http://creativecommons.org/licenses/by-sa/4.0/
P16 How does skin ageing-related basement membrane collagen loss affect the nucleus?
Authors: Simranpreet Kaur Summan, Andrew Mayes, David Gunn, Julien Gautrot, Matthew Caley, Edel O’Toole
Journal: British Journal of Dermatology (2023)

Abstract Skin ageing effects, such as inflammation, oxidative stress, disrupted wound healing and disease, are universal issues. Basement membrane collagens (BMCs) are reduced by ageing and the consequences are poorly understood. BMC sensing of mechanical stress, vital for skin homeostasis, elicits signalling sensed via cytoskeletal actin filaments to the LINC complex (composed of nesprin and sun proteins), nuclear lamina and lamina-associated chromatin. RNAseq data from collagen 7 (Col7) or collagen 17 (Col17) small interfering (si)RNA knockdown primary keratinocytes revealed an opposing enrichment of nuclear function genes. Nesprin-2 was reduced with siCol7 (P &amp;lt; 0.01) vs. siC nontargeting control. We hypothesized that BMC loss in keratinocytes disrupts the nucleocytoskeleton and chromatin organization, dysregulating gene expression and signalling, leading to altered epidermal differentiation and skin barrier impairment. To explore the role of BMC in mechanical stress response, stable short hairpin (sh)RNA knockdowns of Col7 (shCol7) and Col17 (shCol17), and nontargeting control (shC) were generated in immortalized N/TERT cells. Western blot validated knockdowns compared to shC. MTT assays showed a hyperproliferative phenotype with shCol17 compared to shC. Mechanical stretch effect on cells on a flexible membrane was studied by immunofluorescence and Western blot (WB). Images taken from the IN Cell Analyzer confocal microscope were analysed to detect changes in protein expression and localization. Nuclear and cytoplasmic fractions made before and after stretch were analysed by WB. We observed collagen-specific responses to stretch-induced stress with changes to actin and nuclear membrane proteins: emerin and lamin B1, and sun2. Our data are the first to show an altered nucleocytoskeleton in the context of BMC loss and mechanotransduction in skin. This study could provide novel insights into ageing epidermal regulation by BMCs.
Activation of distinct inflammatory pathways in subgroups of LR-MDS
Authors: Marie Schneider, Clara Rolfs, Matthias Trumpp, Susann Winter, Luise Fischer, Mandy Richter, Victoria Menger, Kolja Nenoff, Nora Grieb, Klaus H. Metzeler, Anne Sophie Kubasch, Katja Sockel, Christian Thiede, Jincheng Wu, Janghee Woo, Andreas Brüderle, Lorenz C. Hofbauer, Jörg Lützner, Andreas Roth, Michael Cross, Uwe Platzbecker
Journal: Leukemia (2023)

Abstract Aberrant innate immune signaling has been identified as a potential key driver of the complex pathophysiology of myelodysplastic neoplasms (MDS). This study of a large, clinically and genetically well-characterized cohort of treatment-naïve MDS patients confirms intrinsic activation of inflammatory pathways in general mediated by caspase-1, interleukin (IL)-1β and IL-18 in low-risk (LR)-MDS bone marrow and reveals a previously unrecognized heterogeneity of inflammation between genetically defined LR-MDS subgroups. Principal component analysis resolved two LR-MDS phenotypes with low (cluster 1) and high (cluster 2) levels of IL1B gene expression, respectively. Cluster 1 contained 14/17 SF3B1 -mutated cases, while cluster 2 contained 8/8 del(5q) cases. Targeted gene expression analysis of sorted cell populations showed that the majority of the inflammasome-related genes, including IL1B , were primarily expressed in the monocyte compartment, consistent with a dominant role in determining the inflammatory bone marrow environment. However, the highest levels of IL18 expression were found in hematopoietic stem and progenitor cells (HSPCs). The colony forming activity of healthy donor HSPCs exposed to monocytes from LR-MDS was increased by the IL-1β-neutralizing antibody canakinumab. This work reveals distinct inflammatory profiles in LR-MDS that are of likely relevance to the personalization of emerging anti-inflammatory therapies.
Survival benefit and impact of adjuvant chemotherapy following systemic neoadjuvant chemotherapy in patients with resected pancreas ductal adenocarcinoma: a retrospective cohort study
Authors: Ning Pu, Wenchuan Wu, Siyao Liu, Yuqi Xie, Hanlin Yin, Qiangda Chen, Taochen He, Zhihang Xu, Wenquan Wang, Jun Yu, Liang Liu, Wenhui Lou
Journal: International Journal of Surgery (2023)

Background: Patients with pancreatic ductal adenocarcinoma (PDAC) are increasingly receiving systemic neoadjuvant chemotherapy (NAC), particularly those with borderline resectable and locally advanced disease. However, the specific role of additional adjuvant chemotherapy (AC) in these patients is unknown. The objective of this study is to further assess the clinical benefit and impact of systemic AC in patients with resected PDAC after NAC. Methods: Data on PDAC patients with or without AC following systemic NAC and surgical resection were retrospectively retrieved from the Surveillance, Epidemiology, and End Results (SEER) database between 2006 and 2019. A matched cohort was created using propensity score matching (PSM), and baseline characteristics were balanced to reduce bias. Overall survival (OS) and cancer-specific survival (CSS) were calculated using matching cohorts. Results: The study enrolled a total of 1,589 patients, with 623 (39.2%) in the AC group and 966 (51.8%) in the non-AC group (mean age, 64.0 [9.9] years; 766 [48.2%] were females and 823 [51.8%] were males). All patients received NAC, and among the crude population, 582 (36.6%) received neoadjuvant radiotherapy, while 168 (10.6%) received adjuvant radiotherapy. Following the 1:1 PSM, 597 patients from each group were evaluated further. The AC and non-AC groups had significantly different median OS (30.0 vs. 25.0 mo, P =0.002) and CSS (33.0 vs. 27.0 mo, P =0.004). After multivariate Cox regression analysis, systemic AC was independently associated with improved survival ( P =0.003, HR=0.782; 95%CI, 0.667-0.917 for OS; P =0.004, HR=0.784; 95%CI, 0.663-0.926 for CSS), and age, tumor grade, and AJCC N staging were also independent predictors of survival. Only patients younger than 65 years old and those with a pathological N1 category showed a significant association between systemic AC and improved survival in the subgroup analysis adjusted for these covariates. Conclusion: Systemic AC provides a significant survival benefit in patients with resected PDAC following NAC compared to non-AC patients. Our study discovered that younger patients, patients with aggressive tumors and potentially well response to NAC might benefit from AC to achieve prolonged survival after curative tumor resection.
O06 Exploring host–microbiome–immune interactions using singular and multispecies inoculations in optimized 3D skin models
Authors: Esmé McPolin-Hall, Abish Stephen, Claire  Pardieu, Robert Allaker, Michae  Philpott, Ranjit Bhogal, Jenny Pople, Rosalind  Hannen
Journal: British Journal of Dermatology (2023)

Abstract The human microbiome has a fundamental role in skin homeostasis, barrier function and host immune networks. Specifically, a complex and multispecies microbiome population is required for healthy skin. Acute and chronic skin conditions are often associated with imbalances in host–microbiome–immune interactions. Despite recent advances in skin microbiome research, there are limited models for studying these interactions in healthy and diseased skin; therefore, we have developed a model to investigate how different microbiome profiles alter epidermal homeostasis and T-cell responses. Full-thickness three-dimensional (3D) skin models, developed using primary dermal fibroblasts and keratinocytes isolated from adult skin, were colonized with five human microbiome species: Staphylococcus epidermidis (SE), Staphylococcus capitis (SC), Cutibacterium acnes (CA), Malassezia restricta (MR) and Malassezia globosa (MG), in singular or combined inoculations (5M). Significant changes in epidermal thickness were observed in models colonized with single-species inoculations compared with sterile control models; however, no significant changes in epidermal thickness were observed in models colonised with 5M inoculations. The effect of colonization on epidermal differentiation and barrier formation was observed via the expression of K14, K10, E-cadherin and Ki67. Expression of E-cadherin significantly decreased in models colonized with SC, MR and MG compared with sterile control models. Despite significant changes in epidermal thickness, only MG showed a significant decrease in Ki67 expression compared with sterile control models. No significant changes were observed in K10 and K14 expression following microbial inoculation. Overall, 5M inoculation showed no significant changes in biomarker expression or epidermal thickness compared with sterile control models suggesting a combination of microbes supports epidermal homeostasis as observed in vivo. Commensal microbes have been shown to regulate epidermal barrier function via the aryl hydrocarbon receptor (AhR). Increased nuclear expression of AhR was observed in response to SC, CA and 5M, suggesting these microbes have a greater role in AhR activation. To investigate microbiome–immune interactions in the skin, the expression of CD69, cutaneous lymphocyte antigen (CLA) and HLA-DR on T cells was assessed in response to incubating peripheral blood mononuclear cells with conditioned media from inoculated models. SE caused a significant increase in CD3+/CLA+ cells and 5M significantly reduced CD3+/HLA-DR+ cells. RNA sequencing aims to support current analysis and elucidate any more targets to be explored. Our findings suggest different microbial profiles alter epidermal barrier formation and immune responses.